TY - JOUR
T1 - rpoB gene mutations among Mycobacterium tuberculosis isolates from extrapulmonary sites
AU - Khosravi, Azar Dokht
AU - Meghdadi, Hossein
AU - Ghadiri, Ata A.
AU - Alami, Ameneh
AU - Sina, Amir Hossein
AU - Mirsaeidi, Mehdi
N1 - Funding Information:
This work is part of thesis of Hossein Meghdadi, which has been approved in the Infectious and Tropical Diseases Research Center and was financially supported by a grant (no. A-1209) from Research affairs, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran. Seventy formalin-fixed, paraffin-embedded (FFPE) pathologic blocks (lymph node 25, skin 8, breast 5, colon 4, thyroid 2, soft tissue 4, bladder, omentum, and kidney each one sample) and pleural effusion 19, with histopathological characteristics of granulomatous inflammation and caseous necrosis compatible with EPTB, were collected from the Department of Pathology, University teaching hospital, Ahvaz Jundishapur University of Medical Sciences, Iran.
Publisher Copyright:
© 2018 APMIS. Published by John Wiley & Sons Ltd
PY - 2018/3
Y1 - 2018/3
N2 - The aim of this study was to analyze mutations occurring in the rpoB gene of Mycobacterium tuberculosis (MTB) isolates from clinical samples of extrapulmonary tuberculosis (EPTB). Seventy formalin-fixed, paraffin-embedded samples and fresh tissue samples from confirmed EPTB cases were analyzed. Nested PCR based on the rpoB gene was performed on the extracted DNAs, combined with cloning and subsequent sequencing. Sixty-seven (95.7%) samples were positive for nester PCR. Sequence analysis of the 81 bp region of the rpoB gene demonstrated mutations in 41 (61.2%) of 67 sequenced samples. Several point mutations including deletion mutations at codons 510, 512, 513 and 515, with 45% and 51% of the mutations in codons 512 and 513 respectively were seen, along with 26% replacement mutations at codons 509, 513, 514, 518, 520, 524 and 531. The most common alteration was Gln → His, at codon 513, presented in 30 (75.6%) isolates. This study demonstrated sequence alterations in codon 513 of the 81 bp region of the rpoB gene as the most common mutation occurred in 75.6% of molecularly confirmed rifampin-resistant strains. In addition, simultaneous mutation at codons 512 and 513 was demonstrated in 34.3% of the isolates.
AB - The aim of this study was to analyze mutations occurring in the rpoB gene of Mycobacterium tuberculosis (MTB) isolates from clinical samples of extrapulmonary tuberculosis (EPTB). Seventy formalin-fixed, paraffin-embedded samples and fresh tissue samples from confirmed EPTB cases were analyzed. Nested PCR based on the rpoB gene was performed on the extracted DNAs, combined with cloning and subsequent sequencing. Sixty-seven (95.7%) samples were positive for nester PCR. Sequence analysis of the 81 bp region of the rpoB gene demonstrated mutations in 41 (61.2%) of 67 sequenced samples. Several point mutations including deletion mutations at codons 510, 512, 513 and 515, with 45% and 51% of the mutations in codons 512 and 513 respectively were seen, along with 26% replacement mutations at codons 509, 513, 514, 518, 520, 524 and 531. The most common alteration was Gln → His, at codon 513, presented in 30 (75.6%) isolates. This study demonstrated sequence alterations in codon 513 of the 81 bp region of the rpoB gene as the most common mutation occurred in 75.6% of molecularly confirmed rifampin-resistant strains. In addition, simultaneous mutation at codons 512 and 513 was demonstrated in 34.3% of the isolates.
KW - Extrapulmonary tuberculosis
KW - Mycobacterium tuberculosis
KW - mutation
KW - resistance
KW - rifampin
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U2 - 10.1111/apm.12804
DO - 10.1111/apm.12804
M3 - Article
C2 - 29484749
AN - SCOPUS:85042542966
VL - 126
SP - 241
EP - 247
JO - APMIS
JF - APMIS
SN - 0903-4641
IS - 3
ER -